Vanillic acid retains redox status in hepg2 cells during hyperinsulinemic shock using the mitochondrial pathway

Journal Pre-proofVanillic acid retains redox status in HepG2 cells during hyperinsulinemic shock using the mitochondrial pathway Sreelekshmi Mohan, Genu George, K.G.. & Raghu K.G, Vanill

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Vanillic acid retains redox status in HepG2 cells during hyperinsulinemic shock using

the mitochondrial pathway

Sreelekshmi Mohan, Genu George, K.G Raghu

PII: S2212-4292(21)00141-3

DOI: https://doi.org/10.1016/j.fbio.2021.101016

Reference: FBIO 101016

To appear in: Food Bioscience

Received Date: 15 December 2019

Revised Date: 17 March 2021

Accepted Date: 18 March 2021

Please cite this article as: Mohan S., George G & Raghu K.G, Vanillic acid retains redox status in

HepG2 cells during hyperinsulinemic shock using the mitochondrial pathway, Food Bioscience, https:// doi.org/10.1016/j.fbio.2021.101016

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Author statement

Sreelekshmi Mohan conducted the experiments, collected, analyzed and interpreted the data and she wrote the first draft of the manuscript Dr Genu George edited the manuscript Dr K.G Raghu designed work plan, concept, interpreted the data, contributed intellectual content

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Vanillic acid retains redox status in HepG2 cells during

EDTA ethylenediaminetetraacetic acid

SPSS Statistical Package for the Social Sciences

al., 2015) These effects were reversed using antioxidants such as N-acetyl-cysteine, superoxide

2 Materials and methods

2.2 Establishment of IR through hyperinsulinemic shock of HepG2 cells

2.2.2 Alterations of insulin signaling pathway

The cells after treatment were incubated with DCFH-DA at 37 oC for 20 min Then the cells

buffer (100 μL) and co-substrate mix (50 μL) were added to respective wells Then 20 μL of

EDTA as previously and the cell pellets were sonicated (Elmasonic, Elmasonic S 30 (H), 37 kHz

2.14 Mitochondrial membrane potential (∆ψm)

at 25 V for 15 min to polyvinylidene difluoride (PVDF) membrane (a non-reactive thermoplastic

significantly (p≤0.05) effective in preventing ROS formation by 88.2 and 99.6%, respectively,

prevented the alteration of ∆ψm which was evident from the significant increase (p≤0.05) of red

was reduced (25%) as compared to control Co-treatment with VA at 5 and 10 μM significantly

2017) The efficacy of insulin-stimulated glucose uptake was also reduced with HI (Fig 1a)

One of the risk factors associated with OS is glycation Glycation is one among various

et al., 2015) These results showed that IR induced alterations in redox status of the cell is

the fission rates (Van der Bliek et al., 2013) Hyperinsulinemia results in an overexpression of

important role in the improvement of insulin sensitivity in insulin-resistant individuals (Jeon,

the development of VA-based nutraceuticals All these beneficial properties of VA could be

Choi, J., Chandrasekaran, K., Inoue, T., Muragundla, A., & Russell, J W (2014) PGC-1α

Espinosa-Diez, C., Miguel, V., Mennerich, D., Kietzmann, T., Sánchez-Pérez, P., Cadenas, S., &

Ghosh, A., Bera, S., Ray, S., Banerjee, T., & Ray, M (2011) Methylglyoxal induces

Itoh, A., Isoda, K., Kondoh, M., Kawase, M., Watari, A., Kobayashi, M., Tamesada, M., & Yagi,

Lu, S C (2013) Glutathione synthesis Biochimica et Biophysica Acta (BBA) - General

Michael, M D., Kulkarni, R N., Postic, C., Previs, S F., Shulman, G I., Magnuson, M A., &

Rauscher, F M., Sanders, R A., & Watkins III, J B (2001) Effects of coenzyme Q10 treatment

Song, F., & Schmidt, A M (2012) Glycation and insulin resistance: Novel mechanisms and

Vinoth, A., & Kowsalya, R (2018) Chemopreventive potential of vanillic acid against 7,

Yin, L., Mano, J I., Tanaka, K., Wang, S., Zhang, M., Deng, X., & Zhang, S (2017) High level

(1 mM); NAC-IR + N-acetyl cysteine (1 mM) Values are expressed as mean ± SEM where n=6

were used, IC50 -397 µM/mL Amino guanidine (AG) was used as a reference (IC50 - 500 µM)

Fig 11 Aconitase activity Aconitase enzyme activities in different groups C-control cells;

Fig 14 Level of ATP content in different groups. C-control cells; IR-high insulin treated

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Highlights

● Hyperinsulinemia led to oxidative stress, glycation and mitochondrial dysfunction in HepG2 cells

● Vanillic acid protected cells during hyperinsulinemic shock

● The beneficial action of vanillic acid used the AMPK/Sirt1/PGC-1α activation pathway and its potential antiglycation property

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Conflict of interest

The authors confirm that they have no conflicts of interest with respect to the work described in this manuscript

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